实时荧光定量RT-PCR检测ICP4、IFN-γ和IL-18基因在ILTV感染鸡三叉神经节中的表达  

Real-time RT-PCR for detecting ICP4,IFN-γ and IL-18 gene expression in trigeminal ganglia of ILTV infected chicken

作　　者：木古丽[1,2] 王云峰[1] 石星明[1] 童光志[1] 何来[1] 王玫[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室禽传染病研究室,黑龙江哈尔滨150001 [2]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《中国兽医学报》2008年第10期1150-1153,1166,共5页Chinese Journal of Veterinary Science

摘　　要：将20只4月龄SPF鸡人工感染传染性喉气管炎病毒(ILTV)WG株,于感染后39d翅静脉和肌肉注射地塞米松(DEX),感染后5、9、10、11、12、39d及DEX处理后1、2、3、5、6、7、8d采喉拭子,检测病毒DNA,并于感染前和感染后6、10、20、31d及DEX处理后2、4、7、10、30d每组各取1只鸡颈静脉放血致死后无菌采集三叉神经节,提总RNA后利用real-timeRT-PCR的方法定量检测ICP4、IFN-γ和IL-18特异性mRNA。结果表明,在人工感染后处于潜伏状态的病毒能再被激活,并且ICP4基因在病毒还未被激活时就能低水平表达,这种低水平的立即早期(IE)基因的表达能提供持续的抗原刺激来维持神经节中的炎性细胞分泌IFN-γ和IL-18,以至于维持病毒处于潜伏状态而不致激活后感染其他的健康宿主;DEX处理后2d,IFN-γ的表达达到峰值,对病毒的再激活起抑制作用,尽管DEX处理前后IL-18的表达量没有明显变化,推测当潜伏的病毒一旦被激活时,IFN-γ可协同IL-18起到抑制IL-TV的作用。SPF chickens were infected with ILTV strain WG and intravenously injected with dexamethasone （DEX） at 39 days post infection to initiate reactivation. Infectious virus was detected at 5,9,10, 11,12,39 days post infection and DEX treatment （1,2,3,5.6,7,8 days） by detecting virus genome DNA. Trigeminal ganglion was collected before infection, post infection （ 6,10,20,30 days） and DEX treatment （2,4,7,10,30 days）. Samples were subjected to nucleic acid extraction and processed for real time RT-PCR to quantitatively analysis the expression of ICP4, IFN-γ and IL- 18 transcripts. The results indicate that SPF chickens were infected and reactivated successfully. Low-level of ICP4 gene expressed in the absence of detectable reactivation, this low-level expression of ICP4 gene provides antigenic stimulus to maintain inflammatory cellsresponse and trigger IFN-γ and 1L-18 secretion in ganglia, so this can maintain virus in latent state and not reactivate to infect other healthy host. After DEX treatment,the expression of IFN-γ was at a maximum level on day 2, so it is concluded that IFN-γ function can suppress the reactivation of the ILTV infection. Although there is no obvious difference in IL-18 expression before and after DEX treatment. Thereby we hypothesize that IFN-γ can rapidly suppress ILTV with IL-18 once virus was reactivated.

关 键 词：传染性喉气管炎病毒 地塞米松(DEX) 基因表达定量分析 实时荧光定量RT—PCR 鸡 

分 类 号：S852.659.1[农业科学—基础兽医学]